CN104649930A - Synthesis method and application of a complex [Zn(H2L4)2].(H2O) with anticancer activity - Google Patents

Abstract

The invention discloses a synthesis method and application of a complex [Zn(H2L4)2].(H2O) with anticancer activity. [Zn(H2L4)2].(H2O) has the molecular formula of C22H28Br2N2O7Zn and the molecular weight of 657.65. The synthesis method comprises the steps of (1) mixing 2.01g of analytically pure 5-bromosalicylaldehyde and 15ml of absolute ethyl alcohol, then, adding 1.051g of analytically pure 2-amino-2-methyl-1,3-propanediol and 10ml of absolute ethyl alcohol, heating, carrying out reflux stirring, cooling, crystallizing and standing to obtain H3L4 (H3L4=2-((5-bromo-2-hydroxybenzylidene)amino)-2-methyl-1,3-propanediol; (2) dissolving 0.144g of H3L4 and 0.220g of analytically pure zinc acetate dihydrate into 15-20ml of a mixed solution of absolute ethyl alcohol and self-made distilled water which are in the volume ratio of 5:5; and (3) transferring the mixed solution to a reaction kettle of polytetrafluoroethylene, reacting, cooling, filtering and crystallizing. [Zn(H2L4)2].(H2O) can be applied as an anticancer drug. The synthesis method is simple in process, low in cost, good in repeatability and high in yield; and chemical components are easily controlled.

Description

Title complex with anticancer activity [Zn (H 2l 4) 2]. (H 2o) synthesis and application

Technical field

The present invention relates to a kind of title complex with anticancer activity [Zn (H ₂l ⁴) ₂]. (H ₂o) synthesis and application.

Background technology

Recent study finds that salicylic aldehyde Schiff bases title complex has good antibacterial, anti-inflammatory, the biological activity such as antitumor, the synthesis of salicylic aldehyde presence of Schiff-base complex and bioactive research thereof have become the important topic of pharmaceutical chemistry, biologist's research, for social development and technical progress play the effect become more and more important.Particularly in short supply present of efficient new drug, the medicine of exploitation tool high anti-cancer activity solves one of efficient new drug problem method in short supply.Title complex [Zn (H ₂l ⁴) ₂]. (H ₂o) there is unique biological activity, low toxicity that design and synthesis has the application prospect medicine such as antibacterial, antitumor efficiently can be used as, there is potential use.

Summary of the invention

Object of the present invention is exactly be design and synthesis title complex with anticancer activity [Zn (H ₂l ⁴) ₂]. (H ₂o), solvent structure title complex [Zn (H is utilized ₂l ⁴) ₂]. (H ₂o) and as the application of antitumor drug.

Title complex [Zn (the H that the present invention relates to ₂l ⁴) ₂]. (H ₂o) molecular formula is: C ₂₂h ₂₈br ₂n ₂o ₇zn, molecular weight is: 657.65, and crystal structural data is in table one, and bond distance's bond angle data are in table two.Compound [Zn (H ₂l ⁴) ₂]. (H ₂o) all restraining effect is had to growth of tumour cell such as human hepatoma cell strain (BEL-7404), human liver cancer cell (HepG2), cervical cancer cell (HeLa), human bladder cancer cell (T-24) and normal liver cell's strains (HL-7702).

Table one: [Zn (H ₂l ⁴) ₂]. (H ₂o) crystallographic parameter

Table two: [Zn (H ₂l ⁴) ₂]. (H ₂o) bond distance with bond angle (°)

Zn1–O4	2.016(4)	Zn1–N1	2.101(4)
				Zn1–O1	2.060(4)	Zn1–O5	2.271(4)
Zn1–N2	2.085(4)	Zn1–O2	2.271(4)
				O4–Zn1–O1	99.14(16)	N2–Zn1–O5	76.59(16)
O4–Zn1–N2	91.33(16)	N1–Zn1–O5	87.97(16)
				O1–Zn1–N2	96.17(16)	O4–Zn1–O2	90.66(16)
O4–Zn1–N1	103.13(17)	O1–Zn1–O2	164.45(13)
				O1–Zn1–N1	89.73(15)	N2–Zn1–O2	95.63(16)
N2–Zn1–N1	163.35(18)	N1–Zn1–O2	76.23(15)
				O4–Zn1–O5	166.07(15)	O5–Zn1–O2	83.76(17)
O1–Zn1–O5	89.18(16)

Described [Zn (H ₂l ⁴) ₂]. (H ₂o) synthetic method concrete steps are:

(1) analytically pure for 2.01g 5-bromosalicylaldehyde is placed in there-necked flask, the dehydrated alcohol heated and stirred adding 15ml treats that it dissolves completely, then the analytically pure 2-amino-2-methyl-1 of 1.051g is added, ammediol, add the dehydrated alcohol of 10ml again, heating in water bath, design temperature is 65 DEG C, return stirring 120 minutes.The solution obtained cools under room temperature, crystallization of naturally volatilizing, and leaves standstill after 3 days, obtains yellow bulk crystals H ₃l ⁴(H ₃l ⁴=2-((the bromo-2-phenol methylene of 5-) is amino)-2-methyl isophthalic acid, ammediol).

(2) 0.144 gram of H step (1) synthesized ₃l ⁴15-20 ml volumes is dissolved in than being the dehydrated alcohol of 5:5 and making by oneself in the mixing solutions of distilled water with 0.220 gram of analytical pure two water zinc acetate.

(3) proceed in the reactor of tetrafluoroethylene by the solution obtained by step (2), react 120 hours, be cooled to room temperature at 80-90 DEG C, filter, filtrate is volatilized under being placed in room temperature crystallization naturally, obtains monocrystalline level [Zn (H after 10 days ₂l ⁴) ₂]. (H ₂o) title complex.[Zn (H is measured by single crystal diffractometer ₂l ⁴) ₂]. (H ₂o) structure.

The present invention has that technique is simple, with low cost, chemical composition is easy to control, reproducible, product purity is high and output advantages of higher.

Accompanying drawing explanation

Fig. 1 is part H of the present invention ₃l ⁴enforcement figure.

Fig. 2 is title complex of the present invention [Zn (H ₂l ⁴) ₂]. (H ₂o) enforcement figure.

Fig. 3 is title complex of the present invention [Zn (H ₂l ⁴) ₂]. (H ₂o) structure iron.

Fig. 4 is title complex of the present invention [Zn (H ₂l ⁴) ₂]. (H ₂o) three-dimensional structure accumulation graph.

Embodiment

Embodiment:

Title complex [Zn (H ₂l ⁴) ₂]. (H ₂o) molecular formula is: C ₂₂h ₂₈br ₂n ₂o ₇zn, molecular weight is: 657.65, and crystal structural data is in table one, and bond distance's bond angle data are in table two.

Title complex [Zn (H ₂l ⁴) ₂]. (H ₂o) synthetic method concrete steps are:

(1) analytically pure for 2.01g 5-bromosalicylaldehyde is placed in there-necked flask, the dehydrated alcohol heated and stirred adding 15ml treats that it dissolves completely, then the analytically pure 2-amino-2-methyl-1 of 1.051g is added, ammediol, add the dehydrated alcohol of 10ml again, heating in water bath, design temperature is 65 DEG C, return stirring about 120 minutes; The solution obtained cools under room temperature, crystallization of naturally volatilizing, and leaves standstill after 3 days, obtains yellow bulk crystals H ₃l ⁴(H ₃l ⁴=2-((the bromo-2-phenol methylene of 5-) is amino)-2-methyl isophthalic acid, ammediol).

(2) 0.144 gram of H step (1) synthesized ₃l ⁴15-20 ml volumes is dissolved in than being the dehydrated alcohol of 5:5 and making by oneself in the mixing solutions of distilled water with 0.220 gram of analytical pure two water zinc acetate.

(3) proceed in the reactor of tetrafluoroethylene by the solution obtained by step (2), react 120 hours, be cooled to room temperature at 80-90 DEG C, filter, filtrate is volatilized under being placed in room temperature crystallization naturally, obtains monocrystalline level [Zn (H after 10 days ₂l ⁴) ₂]. (H ₂o) title complex.[Zn (H is measured by single crystal diffractometer ₂l ⁴) ₂]. (H ₂o) structure.

[Zn (H with anticancer activity ₂l ⁴) ₂]. (H ₂o) proliferation inhibition activity of various human tumor cell line is tested:

(1) cell strain and cell cultures:

Human hepatoma cell strain (BEL-7404) is selected in this experiment, human liver cancer cell (HepG2), cervical cancer cell (HeLa), human bladder cancer cell (T-24), 5 kinds of human cell lines such as normal liver cell's strain (HL-7702).All cells strain is all cultivated in the RPMI-1640 nutrient solution containing 10wt% calf serum, 100U/mL penicillin, 100U/mL Streptomycin sulphate, puts 37 DEG C containing volumetric concentration 5%CO ₂cultivate in incubator.

(2) preparation of testing compound:

[Zn (H used ₂l ⁴) ₂]. (H ₂o) purity>=95%, be mixed with the whole solution of 20 μm of ol/L after being diluted by its DMSO liquid storage physiological buffer, wherein final concentration≤1% of solubility promoter DMSO, under testing this concentration, compound is to the suppression degree of various growth of tumour cell.

(3) cell growth inhibition test (mtt assay):

1) tumour cell of taking the logarithm vegetative period, after tryptic digestion, the cell suspension that concentration is 5000/ml is mixed with the nutrient solution containing 10% calf serum, be inoculated in 96 well culture plates with every hole 190 μ l, make hole, cell density to 1000 ~ 10000 to be measured (the aseptic PBS of marginal pore fills);

2) 5%CO ₂, hatch 24 hours for 37 DEG C, be paved with at the bottom of hole to cell monolayer, every hole adds the medicine 10 μ L of finite concentration gradient, and each concentration gradient establishes 4 multiple holes;

3) 5%CO ₂, hatch 48 hours for 37 DEG C, observe under inverted microscope;

4) every hole adds the MTT solution (5mg/mL PBS, i.e. 0.5%MTT) of 10 μ L, continues cultivation 4 hours;

5) stop cultivating, carefully suck nutrient solution in hole, the DMSO that every hole adds 150 μ L fully dissolves first a ceremonial jade-ladle, used in libation precipitation, and after vibrator mixing, be 570nm at microplate reader wavelength, reference wavelength is the optical density value that 450nm measures each hole;

6) zeroing hole (substratum, MTT, DMSO) is set simultaneously, control wells (the medicine dissolution medium of cell, same concentrations, nutrient solution, MTT, DMSO).

7) according to the optical density value (OD value) recorded, judge viable cell quantity, OD value is larger, and cytoactive is stronger.Utilize formula:

Calculate the inhibiting rate of drug on tumor Growth of Cells, then calculate the IC of each test-compound to various human tumor cell line and Human normal hepatocyte strain respectively with Bliss method ₅₀value.

Title complex [Zn (H ₂l ⁴) ₂]. (H ₂o) to the inhibiting rate of human hepatoma cell strain (BEL-7404), human liver cancer cell (HepG2), cervical cancer cell (HeLa), human bladder cancer cell (T-24) and normal liver cell's strain (HL-7702) growth of tumour cell in table three, to the IC of various human tumor cell line and Human normal hepatocyte strain ₅₀value is in table four.

Table three: [Zn (H ₂l ⁴) ₂]. (H ₂o) to the inhibiting rate of different cell strain

BEL-7404	HepG2	HeLa	T-24	HL-7702
					43.77±1.12	23.69±1.14	45.03±0.88	49.7±1.54	38.09±1.73

Table four: [Zn (H ₂l ⁴) ₂]. (H ₂o) to the half-inhibition concentration (IC of different cell strain ₅₀, μM)

BEL-7404	HepG2	HeLa	T-24	HL-7702
					21.35±1.38	33.15±1.28	20.11±0.69	17.06±1.81	26.66±1.27

Claims (2)

1. a title complex with anticancer activity [Zn (H ₂l ⁴) ₂]. (H ₂o), it is characterized in that title complex [Zn (H ₂l ⁴) ₂]. (H ₂o) molecular formula is: C ₂₂h ₂₈br ₂n ₂o ₇zn, molecular weight is: 657.65, and crystal structural data is in table one, and bond distance's bond angle data are in table two;

Described title complex [Zn (H ₂l ⁴) ₂]. (H ₂o) synthetic method concrete steps are:

(1) analytically pure for 2.01g 5-bromosalicylaldehyde is placed in there-necked flask, the dehydrated alcohol heated and stirred adding 15ml treats that it dissolves completely, then the analytically pure 2-amino-2-methyl-1 of 1.051g is added, ammediol, add the dehydrated alcohol of 10ml again, heating in water bath, design temperature is 65 DEG C, return stirring 120 minutes; The solution obtained cools under room temperature, crystallization of naturally volatilizing, and leaves standstill after 3 days, obtains yellow bulk crystals H ₃l ⁴, H ₃l ⁴for 2-((the bromo-2-phenol methylene of 5-) is amino)-2-methyl isophthalic acid, ammediol;

(2) by H that 0.144 gram of step (1) is synthesized ₃l ⁴15-20 ml volumes is dissolved in than being the dehydrated alcohol of 5:5 and making by oneself in the mixing solutions of distilled water with 0.220 gram of analytical pure two water zinc acetate;

(3) solution of step (2) gained is proceeded in the reactor of tetrafluoroethylene, react 120 hours at 80-90 DEG C, be cooled to room temperature, filter, filtrate is volatilized under being placed in room temperature crystallization naturally, obtains monocrystalline level title complex [Zn (H after 10 days ₂l ⁴) ₂]. (H ₂o); Title complex [Zn (H is measured by single crystal diffractometer ₂l ⁴) ₂]. (H ₂o) structure;

Table one: { [Zn (H ₂l ⁴) ₂]. (H ₂o) crystallographic parameter }

Table two: { [Zn (H ₂l ⁴) ₂]. (H ₂o) bond distance } with bond angle (°)

Zn1–O4 2.016(4) Zn1–N1 2.101(4) Zn1–O1 2.060(4) Zn1–O5 2.271(4) Zn1–N2 2.085(4) Zn1–O2 2.271(4) O4–Zn1–O1 99.14(16) N2–Zn1–O5 76.59(16) O4–Zn1–N2 91.33(16) N1–Zn1–O5 87.97(16) O1–Zn1–N2 96.17(16) O4–Zn1–O2 90.66(16) O4–Zn1–N1 103.13(17) O1–Zn1–O2 164.45(13) O1–Zn1–N1 89.73(15) N2–Zn1–O2 95.63(16) N2–Zn1–N1 163.35(18) N1–Zn1–O2 76.23(15) O4–Zn1–O5 166.07(15) O5–Zn1–O2 83.76(17) O1–Zn1–O5 89.18(16)

。

2. title complex according to claim 1 [Zn (H ₂l ⁴) ₂]. (H ₂o) application, is characterized in that title complex [Zn (H ₂l ⁴) ₂]. (H ₂o) apply as antitumor drug.